Part:BBa_K4761100

pSA expression vector

pSA plasmid vector. Expression vector for rhodioloside production derived from pSC101 vector. It has Cm resistance and low copy number(up to 5 copies per cell). BamHI and KpnI are necessary for gene installation.

pSC101 is a narrow host range, low-copy plasmid from Salmonella commonly used for genetically manipulating Escherichia coli, and it was the first plasmid vector used for cloning.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 2920
  Illegal EcoRI site found at 3420
  Illegal SpeI site found at 1669
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 2920
  Illegal EcoRI site found at 3420
  Illegal SpeI site found at 1669
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 2920
  Illegal EcoRI site found at 3420
  Illegal BglII site found at 1687
  Illegal BamHI site found at 2
  Illegal XhoI site found at 3340
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 2920
  Illegal EcoRI site found at 3420
  Illegal SpeI site found at 1669
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 2920
  Illegal EcoRI site found at 3420
  Illegal SpeI site found at 1669
  Illegal NgoMIV site found at 217
  
• 1000
  COMPATIBLE WITH RFC[1000]

Characterization

In 2023 HiZJU-China, we used pSA plasmid vector for tyrosol production module on rhodioloside production.

Fig.1 Verification for original plasmids backbones and genes. pSA backbone is marked out.

Fermentation strain pSA-ARO10-adh6 + pET-UGT85A1, known as original fermentation strain type 2, is constructed to evaluate its performance in rhodioloside production. It has a yield of over 7mg/L 36h after induction via IPTG, proving its capability to express the tyrosol generation unit(BBa_K4761200).

Fig.2 HPLC analysis for fermentation strain pSA-ARO10-adh6 + pET-UGT85A1. Rhodioloside peak marked out.